Synthesis, in-vitro cytotoxicity of 4H-benzo[h]chromene derivatives and structure-activity relationships of 4-aryl group and 3-, 7-positions

Article abstract of DOI:10.1515/chempap-2016-0049

A series of 2-amino-4H-benzo[h]chromene and 2,7-diamino-4H-benzo[h]chromene derivatives were prepared as potential cytotoxic agents. The structures of the synthesised compounds were established on the basis of spectral data. The in-vitro cytotoxic activit

Full text of DOI:10.1515/chempap-2016-0049

Chemical Papers 70 (9) 1279–1292 (2016)
DOI: 10.1515/chempap-2016-0049
ORIGINAL PAPER
Synthesis, in-vitro cytotoxicity of 4H-benzo[ ]chromene derivatives
h
and structure–activity relationships of 4-aryl group
and 3-, 7-positions
b
c
aAhmed M. El-Agrody*, Heba K. Abd El-Mawgoud, Ahmed M. Fouda,
aEssam S. A. E. H. Khattab
aChemistry Department, Faculty of Science, Al-Azhar University, Nasr City 11884, Cairo, Egypt
bChemistry Department, College of Women for Arts, Science and Education, Ain Shams University,
Heliopolis 11757, Cairo, Egypt
cChemistry Department, Faculty of Science, King Khalid University, Abha, 61413, P.O. Box 9004, Saudi Arabia
Received 13 August 2015; Revised 26 January 2016; Accepted 29 January 2016
A series of 2-amino-4H-benzo[h]chromene and 2,7-diamino-4H-benzo[h]chromene derivatives were
prepared as potential cytotoxic agents. The structures of the synthesised compounds were estab-
lished on the basis of spectral data. The in-vitro cytotoxic activity of the synthesised compounds
against the cell lines MCF-7, HCT-116 and HepG-2 was investigated in comparison with vinblas-
tine and colchicine, using an MTT colorimetric assay. The structure–activity relationship of 4H-
benzo[h]chromenes with modification at the 3-, 4- and 7-positions was explored. The results of the
anti-tumour evaluation revealed that compounds VIIIc, VIId, VIIb, VIIe, VIIIg and VIIIc, VIId,
VIIb, VIIe, VIIIg, VIIc, VIIIe, Vf, IIIa inhibited the growth of MCF-7 in comparison with vinblas-
tine and colchicine, while VIIb, VIId, VIIe, IIIa, VIIa, VIIIc, VIIc, IIId, IIIg, IIIf, IIIb, IIIh, VIIIb,
VIIIa, VIIIe, IIIc, Vg, IIIe, VIIIg, Vf, IIIf inhibited the growth of HCT-116 in comparison with
colchicine. In addition, compounds VIIe, IIIg, IIIa, VIIc and VIIe, IIIg, IIIa, VIIc, VIIb, VIIa, VIIIf,
VIIIe inhibited the growth of HepG-2 in comparison with vinblastine and colchicine, respectively.
c
ꢀ 2016 Institute of Chemistry, Slovak Academy of Sciences
Keywords: 1-naphthol, 5-amino-1-naphthol, 4H-benzo[h]chromenes, cytotoxicity, SAR
Introduction
were also active in drug-resistant cancer cell-lines in-
cluding the vascular-disrupting, paclitaxel-resistant,
multi-drug resistant tumour cells, and were found to
be highly active in several anticancer animal mod-
els (Gourdeau et al., 2004; Kasibhatla et al., 2004;
Endo et al., 2010). On the other hand, a diverse
group of 4H-chromene compounds with a substituted
or oligo-substituted phenyl ring at the 4-position and
other groups at the 7-position have been reported
as cytotoxic and anticancer agents (Rampa et al.,
2005; Sabry et al., 2011; El-Agrody et al., 2014a,
2014b). Fused chromene ring systems have blood
Chromene-based compounds have been reported
as possessing many pharmacological activities and
antimicrobial properties (Alvey et al., 2009; Kumar
et al., 2009; Raj et al., 2009; Kidwai et al., 2010;
Li et al., 2010; Liu et al., 2010); however, recent
reports have demonstrated the potential of 4-aryl-
4H-chromenes as apoptosis-inducers (Kemnitzer et
al., 2007, 2008; Mahmoodi et al., 2010). These com-
pounds were found to be tubulin destabilisers, bind-
ing at or close to the binding site of colchicine. They
*Corresponding author, e-mail: elagrody am@yahoo.com

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A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
Table 1. Characterisation data of compounds IIIa–IIIh
Compound
Formula
Mr
Colour
Yield/%
M.p./◦C
IIIa
IIIb
IIIc
IIId
IIIe
IIIf
C
20H14N2O
298.34
316.33
332.78
377.23
312.36
328.36
343.34
383.44
Colourless
Yellow
Yellow
Colourless
Yellow
Yellow
85
86
88
90
90
88
88
82
217, 218a
233, 232b
234, 231b
240
206, 204b
196, 195a
241, 240a
222
C20H13FN2O
C20H13ClN2O
C20H13BrN2O
C21H16N2O
C21H16N2O2
C20H13N3O3
C24H21N3O2
IIIg
IIIh
Yellow
Yellow
a) According to Zhang et al. (2007); b) according to Gong et al. (2008).
platelet anti-aggregating effects (Lee et al., 2006), ex-
hibit analgesic activities (Ali & Ibrahim, 2010; Keri
et al., 2010), hypolipidemic activity (Sashidhara et
al., 2011), DNA-breaking activities and mutagenic-
ity (Hiramoto et al., 1997), and are applicable in
the treatment of Alzheimer’s disease (Bru¨hlmann et
al., 2010) and Schizophrenia disorder (Kesten et al.,
1997). Accordingly, it was decided to synthesise some
oligo-substituted 4H-benzo[h]chromenes as potential
cytotoxic agents. Hence, below is described the syn-
thesis of some 4H-benzo[h]chromene derivatives and
their in-vitro cytotoxicity against a variety of human
cancer cell lines. The chemical structures of the com-
pounds studied and their structure-activity relation-
ships (SAR) at the 3-, 4- and 7-positions are discussed
in this work.
benzo[h]chromene-3-carbonitrile (Va–Vh)
derivatives
A solution of 1-naphthol (I) or 5-amino-1-naphthol
(IV) (0.01 mol) in EtOH (30 mL) and piperidine
(0.5 mL) was treated with α-cyano-p-monosubstituted
cinnamonitriles (IIa–IIh) (0.01 mol). The reaction
mixture was heated under reflux for 1 h. The solid
product thus formed was collected by filtration,
washed with MeOH, re-crystallised from ethanol or
benzene; the colours and yield are reported after crys-
tallisation. The composition, properties and spectral
data of the corresponding products III and V are given
in Tables 1–4.
General procedure for preparation of ethyl
2-amino-4-aryl-4H-benzo[h]chromene-3-
carboxylate (VIIa–VIIg)and ethyl 4-aryl-2,7-
diamino-4H-benzo[h]chromene-3-carboxylate
(VIIIa–VIIIg)derivatives
Experimental
Commercial-grade solvents and reagents were pur-
chased from Sigma–Aldrich (St. Louis, MO, USA) and
used without further purification. Melting points were
measured with a Stuart Scientific (UK) apparatus,
and are uncorrected. IR spectra were determined as
KBr pellets on a Jasco FT/IR 460 plus spectropho-
tometer (Jasco, Japan). 1H NMR and 13C NMR spec-
tra were recorded using a Bruker AV 500 MHz spec-
trometer (Bruker, USA). 13C NMR spectra were ob-
tained using distortion-free enhancement by polarisa-
tion transfer (DEPT), where the signals of the CH and
CH3 carbon atoms appear normal (up) and the signals
of the carbon atoms in CH2 environments appear neg-
ative (down). 13C NMR spectra were obtained using
the attached proton test (APT); with this technique,
the signals of the CH and CH3 carbon atoms appears
normal (up) and the signal of the CH2 and Cq en-
vironments appears negative (down). Chemical shifts
(δ) are expressed in parts per million (ppm), and the
coupling constants (J) are reported in Hz. The MS
were measured using a Shimadzu GC/MS-QP5050A
spectrometer (Shimadzu, Japan).
A solution of 1-naphthol (I) or 5-amino-1-naphthol
(IV ) (0.01 mol) in EtOH (30 mL) and piperi-
dine (0.5 mL) was treated with ethyl α-cyano-p-
monosubstitutedcinnamates (VIa–VIg) (0.01 mol).
The reaction mixture was heated under reflux for 2 h.
The solid product thus formed was collected by fil-
tration, washed with MeOH and re-crystallised from
ethanol or benzene; the colours and yield are reported
after crystallisation. The composition, properties and
spectral data of the corresponding products VII and
VIII are given in Tables 5–8.
Anti-tumour screening
Cell culture and cytotoxicity evaluation using via-
bility assay: the target compounds were initially evalu-
ated for in-vitro anti-tumour activity against three dif-
ferent human cell lines: MCF-7, HCT-116 and HepG-
2 (National Cancer Institute, Cairo, Egypt) in com-
parison with vinblastine and colchicine. The measure-
ments of cell growth, the viabilities and in-vitro cy-
totoxicity evaluation using the viability assay were
determined as described in the literature (Mossman,
1983; Rahman et al., 2001) and the results are listed
General procedure for preparation of 2-amino-
4-aryl-4H-benzo[h]chromene-3-carbonitrile
(IIIa–IIIh) and 4-aryl-2,7-diamino-4H-

A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
Table 2. Spectral data of compounds IIIa–IIIh
1281
Compound
Spectral data
IIIa
IR, ν˜/cm−1: 3447, 3304, 3189 (NH2), 3055, 3021, 2885 (CH), 2204 (CN)
1H NMR (DMSO-d6), δ: 8.28–7.11 (m, 11H, 3Ph), 7.20 (bs, 2H, NH2, D2O exchangeable), 4.91 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.15 (C), 145.67 (C), 142.72 (C), 132.67 (C), 128.67 (CH), 128.29 (CH), 127.64 (CH),
126.89 (CH), 126.73 (CH), 126.64 (CH), 126.20 (CH), 123.87 (CH), 122.74 (C), 120.68 (CH), 120.49 (C), 117.91
(CN), 56.26 (C), 40.91 (CH)
IIIb
IIIc
IIId
IR, ν˜/cm−1: 3459, 3330, 3196 (NH2), 3091, 3034, 2989, 2861 (CH), 2193 (CN)
1H NMR (DMSO-d6), δ: 8.27–7.10 (m, 10H, 3Ph), 7.20 (bs, 2H, NH2, D2O exchangeable), 4.96 (s, 1H,pyran ring)
13C NMR (DMSO-d6), δ: 162.06 (C), 160.12 (C), 142.70 (C), 141.90 (C), 132.69 (C), 129.58 (CH), 129.52 (CH),
128.28 (CH), 127.65 (CH), 126.79 (CH), 126.10 (CH), 123.94 (CH), 122.73 (C), 120.69 (C), 120.38 (CH), 117.69
(CN), 115.49 (CH), 115.32 (CH), 56.17 (C), 40.00 (CH)
IR, ν˜/cm−1: 3453, 3334, 3197 (NH2), 3059, 3042, 2869 (CH), 2192 (CN)
1H NMR (DMSO-d6), δ: 8.29–7.10 (m, 10H, 3Ph), 7.22 (bs, 2H, NH2, D2O exchangeable), 4.96 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.14 (C), 144.63 (C), 142.73 (C), 132.73 (C), 131.52 (C), 129.54 (CH), 128.67 (CH),
127.67 (CH), 126.85 (CH), 126.71 (CH), 126.04 (CH), 123.99 (CH), 122.71 (C), 120.69 (C), 120.32 (CH), 117.38
(CN), 55.85 (C), 40.16 (CH)
IR, ν˜/cm−1: 3458, 3342, 3202 (NH2), 3060, 3031, 2865 (CH), 2190 (CN)
1H NMR (DMSO-d6), δ: 8.27–7.09 (m, 10H, 3Ph), 7.24 (bs, 2H, NH2, D2O exchangeable), 4.95 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.14 (C), 145.04 (C), 142.75 (C), 132.74 (C), 131.58 (CH), 129.92 (CH), 128.28 (CH),
127.66 (CH), 126.84 (CH), 126.70 (C), 126.03 (CH), 123.99 (CH), 122.72 (C), 120.70 (C), 120.33 (CH), 120.06 (C),
117.29 (CN), 55.80 (C), 40.27 (CH)
MS, m/z (Ir/%): 378 (20.02) (M++ 2), 376 (20.35) (M+) with a base peak at 221 (100)
IR, ν˜/cm−1: 3412, 3323, 3203 (NH2), 3057, 3031, 2957, 2837 (CH), 2194 (CN)
IIIe
IIIf
1H NMR (DMSO-d6), δ: 8.27–6.87 (m, 10H, 3Ph), 7.14 (bs, 2H, NH2, D2O exchangeable), 4.85 (s, 1H, pyran ring),
2.28 (s, 3H, CH3)
13C NMR (DMSO-d6), δ: 159.97 (C), 158.15 (C), 142.57 (C), 137.81(C), 132.61 (C), 128.71 (CH), 128.29 (CH),
127.63 (CH), 126.67 (CH), 126.60 (CH), 126.25 (CH), 123.77 (CH), 122.74 (C), 120.67 (C), 120.53 (CH), 118.19
(CN), 55.00 (C), 40.09 (CH), 22.44 (CH3)
IR, ν˜/cm−1: 3452, 3339, 3211 (NH2), 3057, 2932, 2836 (CH), 2190 (CN)
1H NMR (DMSO-d6), δ: 8.26–6.87 (m, 10H, 3Ph), 7.12 (bs, 2H, NH2, D2O exchangeable), 4.85 (s, 1H, pyran ring),
3.72 (s, 3H, OCH3)
13C NMR (DMSO-d6), δ: 159.96 (C), 158.15 (C), 142.57 (C), 137.81(C), 132.61 (C), 128.70 (CH), 128.29 (CH),
127.63 (CH), 126.68 (CH), 126.60 (CH), 123.78 (CH), 122.73 (C), 120.66 (C), 120.52 (CH), 118.20 (CN), 114.02
(CH), 56.55 (CH3), 55.01 (C), 40.07 (CH)
MS, m/z (Ir/%): 362 (4.31) (M+) with a base peak at 75 (100)
IR, ν˜/cm−1: 3460, 3359, 3210 (NH2), 3070, 2953, 2864 (CH), 2187 (CN)
IIIg
IIIh
1H NMR (DMSO-d6), δ: 8.21–7.12 (m, 10H, 3Ph), 7.37 (bs, 2H, NH2, D2O exchangeable), 5.18 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.32 (C), 152.93 (C), 146.47 (C), 142.93 (C), 132.88 (C), 128.99 (CH), 127.69 (CH),
127.00 (CH), 126.80 (CH), 125.88 (CH), 124.18 (CH), 124.03 (CH), 122.73 (C), 120.73 (C), 120.14 (CH), 116.57
(CN), 55.21 (C), 40.68 (CH)
IR, ν˜/cm−1: 3401, 3329, 3206 (NH2), 3055, 3016, 2968, 2938 (CH), 2192 (CN)
1H NMR (DMSO-d6), δ: 8.25–6.87 (m, 10H, 3Ph), 7.10 (bs, 2H, NH2), 4.79 (s, 1H, pyran ring), 3.71–3.69 (m, 4H,
2CH2), 3.06–3.04 (m, 4H, 2CH2)
13C NMR (DMSO-d6), δ: 159.87 (C), 149.84 (C), 142.46 (C), 136.34 (C), 132.50 (C), 128.22 (CH), 128.11 (CH),
127.55 (CH), 126.56 (CH), 126.50 (CH), 123.65 (CH), 122.65 (C), 120.56 (CH), 120.51 (C), 118.27 (CN), 115.16
(CH), 65.99 (CH2), 56.53 (C), 48.32 (CH2), 39.90 (CH)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.22 (CH ↑), 128.11 (CH ↑), 127.55 (CH ↑), 126.56 (CH
↑), 126.50 (CH), 123.65 (CH ↑), 120.56 (CH ↑), 115.16 (CH ↑) 65.99 (CH2 ↓), 48.32 (CH2 ↓), 39.90 (CH ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.22 (CH ↑), 128.11 (CH ↑), 127.55 (CH ↑), 126.56 (CH ↑),
126.50 (CH), 123.65 (CH ↑), 120.56 (CH ↑), 115.16 (CH ↑), 39.90 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.22 (CH ↑), 128.11 (CH ↑), 127.55 (CH ↑), 126.56
(CH ↑), 126.50 (CH), 123.65 (CH ↑), 120.56 (CH ↑), 115.16 (CH ↑), 65.99 (CH2 ↑), 48.32 (CH2 ↑), 39.90 (CH ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 159.87 (C ↓), 149.84 (C ↓), 142.46 (C ↓), 136.34 (C ↓), 132.50
(C ↓), 128.22 (CH ↑), 128.11 (CH ↑), 127.55 (CH ↑), 126.56 (CH ↑), 126.50 (CH ↑), 123.65 (CH ↑), 122.65 (C ↓),
121.56 (C ↓), 120.51 (CH ↑), 118.27 (CN ↓), 115.16 (CH ↑), 65.99 (CH2 ↓), 56.53 (C ↓), 48.32 (CH2 ↓), 39.90 (CH
↑)
MS, m/z (Ir/%): 383 (49.34) (M+) with a base peak at 221 (100)
in Table 9 and Fig. 5.
Results and discussion
The treatment of 1-naphthol (I) with α-cyano-p-
monosubstituted cinnamonitriles (IIa–IIh) in ethano-
lic piperidine under reflux gave the corresponding 2-
amino-4-aryl-4H-benzo[h]-chromene-3-carbonitrile
(IIIa–IIIh) derivatives shown in Fig. 1. The composi-
tion, properties and spectral data of the corresponding

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A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
Table 3. Characterisation data of prepared compounds Va–Vh
Compound
Formula
Mr
Colour
Yield/%
M.p./◦C
Va
Vb
Vc
Vd
Ve
Vf
C
20H15N3O
313.35
331.34
347.8
392.25
327.38
343.38
358.35
398.46
Grey
Grey
Grey
Grey
Grey
Grey
Red
85
87
81
80
83
88
79
78
267, 267a
235
C20H14FN3O
C20H14ClN3O
C20H14BrN3O
C21H17N3O
C21H17N3O2
C20H14N4O3
C24H22N4O2
266, 267a
226
230
269, 267a
239
Vg
Vh
Red
235
a) According to Abd-El-Aziz et al. (2004).
Fig. 1. Synthesis of 2-amino-4H-benzo[h]chromene derivatives (IIIa–IIIh).
Fig. 2. Synthesis of 2,7-diamino-4H-benzo[h]chromene derivatives (Va–Vh).
Fig. 3. Synthesis of ethyl 2-amino-4H-benzo[h]chromene-3-carboxylate derivatives (VIIa–VIIg).
products III are summarised in Tables 1 and 2.
In a similar manner, the treatment of 5-amino-1-
naphthol (IV) with α-cyano-p-mono-substituted cin-
namonitriles (IIa–IIh) in ethanolic piperidine under
reflux gave the corresponding 4-aryl-2,7-diamino-4H-
benzo[h]chromene-3-carbonitrile (Va–Vh) derivatives
shown in Fig. 2. The composition, properties and spec-
tral data of the corresponding products V are sum-
marised in Tables 3 and 4.
sis of IR, 1H NMR, 13C NMR, 13C NMR-DEPT,
13C NMR-APT and MS data. The IR spectra of
IIIa–IIIh and Va–Vh showed the appearance of −a1n
NH2 stretch at 3460–3401 cm−1, 3359–3304 cm
,
3211–3189 cm−1 a CN stretch at 2204–2187 cm−1
for IIIa–IIIh and an NH2 stretch at 3469–3422 cm−1
,
,
3399–3336 cm−1, 3332–3309 cm−1, 3297–3257 cm−1
3209–3167 cm−1 a CN stretch at 2200–2184 cm−1
13
for Va–Vh. The 1H and
C NMR spectra of IIIa–
Structures III and V were established on the ba-
IIIh and Va–Vh revealed the presence of 4H sig-

A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
1283
nals at δ of 5.18–4.73 (s, 1H, H-4) and 40.91–
39.90 (C-4). The 13C NMR-DEPT spectra at 45◦,
90◦, 135◦, 13C NMR-APT spectra and the mass
spectra of compounds III and V provided addi-
tional evidence in support of the proposed struc-
tures.
forded ethyl 2-amino-4-aryl-4H-benzo[h]chromene-3-
carboxylate (VIIa–VIIg) derivatives (Fig. 3).The com-
position, properties and spectral data of the corre-
sponding products VII are summarised in Tables 5
and 6.
In a similar manner, the reaction of 5-amino-1-
naphthol (IV) with ethyl α-cyano-p-monosubstituted
cinnamates (VIa–VIg) afforded the ethyl 4-aryl-2,7-
The interaction of 1-naphthol (I) with ethyl
α-cyano-p-monosubstitutedcinnamates (VIa–VIg) af-
Table 4. Spectral data of compounds Va–Vh
Compound
Spectral data
Va
IR, ν˜/cm−1: 3424, 3336, 3325, 3297, 3175 (2 NH2), 3023, 3001 (CH), 2200 (CN)
1H NMR (DMSO-d6), δ: 7.75–6.70 (m, 10H, 3Ph), 7.10 (bs, 2H, NH2-2, D2O exchangeable), 5.78 (bs, 2H, NH2-7,
D2O exchangeable), 4.84 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.32 (C), 145.85 (C), 144.88 (C), 142.86 (C), 128.62 (CH), 128.29 (CH), 127.56 (CH),
127.44 (C), 124.01 (CH), 123.34 (CH), 122.01 (C), 120.63 (C), 118.62 (CH), 117.42 (CN), 108.36 (CH), 107.96 (CH),
56.09 (C), 40.88 (CH)
Vb
IR, ν˜/cm−1: 3450, 3386, 3332, 3257, 3203 (2 NH2), 3028, 3006 (CH), 2195 (CN)
1H NMR (DMSO-d6), δ: 7.76–6.71 (m, 9H, 3Ph), 7.13 (bs, 2H, NH2-2, D2O exchangeable), 5.80 (bs, 2H, NH2-7,
D2O exchangeable), 4.89 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 161.93 (C), 160.19 (C), 144.82 (C), 142.75 (C), 141.98 (C), 129.41 (CH), 128.21 (CH),
127.41 (C), 123.94 (CH), 123.16 (C), 120.47 (C), 118.63 (CH), 117.13 (CN), 115.36 (CH), 108.34 (CH), 107.89 (CH),
55.93 (C), 39.98 (CH)
MS, m/z (Ir/%): 331 (23.84) (M+) with a base peak at 75 (100)
Vc
Vd
IR, ν˜/cm−1: 3463, 3341, 3311, 3287, 3197 (2 NH2), 3021, 3016 (CH), 2188 (CN)
1H NMR (DMSO-d6), δ: 7.76–6.74 (m, 9H, 3Ph), 7.15 (bs, 2H, NH2-2, D2O exchangeable), 5.75 (bs, 2H, NH2-7,
D2O exchangeable), 4.90 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.23 (C), 144.83 (C), 144.74 (C), 142.79 (C), 131.32 (C), 129.38 (CH), 128.52 (CH),
128.21 (CH), 127.44 (C), 123.92 (CH), 123.09 (C), 120.40 (C), 118.68 (CH), 116.79 (CN), 108.35 (CH), 107.85 (CH),
55.61 (C), 40.08 (CH)
IR, ν˜/cm−1: 3469, 3364, 3309, 3293, 3167 (2 NH2), 3011, 3019 (CH), 2186 (CN)
1H NMR (DMSO-d6), δ: 7.78–6.74 (m, 9H, 3Ph), 7.17 (bs, 2H, NH2-2, D2O exchangeable), 5.81 (bs, 2H, NH2-7,
D2O exchangeable), 4.89 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.23 (C), 145.15 (C), 144.82 (C), 142.81 (C), 131.54 (CH), 129.77 (CH), 128.21 (CH),
127.45 (C), 123.93 (CH), 123.11 (C), 120.42 (C), 119.87 (C)118.69 (CH), 116.72 (CN), 108.40 (CH), 107.90 (CH),
55.58 (C), 40.19 (CH)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 131.54 (CH ↑), 129.77, (CH ↑), 128.21 (CH ↑), 123.93 (CH
↑), 118.69 (CH ↑), 108.40 (CH ↑), 107.90 (CH ↑), 40.19 (CH ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 131.54 (CH↑), 129.77, (CH ↑), 128.21 (CH ↑), 123.93 (CH ↑),
118.69 (CH ↑), 108.40 (CH ↑), 107.90 (CH ↑), 40.19 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 131.54 (CH ↑), 129.77, (CH ↑), 128.21 (CH ↑),
123.93 (CH ↑), 118.69 (CH ↑), 108.40 (CH ↑), 107.90 (CH ↑), 40.19 (CH ↑)
13CNMR-APT spectrum CH, CH3 (up), CH2, Cq (down), δ: 160.23 (C ↓), 145.15 (C↓), 144.82 (C ↓), 142.81 (C ↓),
131.54 (CH ↑), 129.77 (CH ↑), 128.21 (CH ↑), 127.45 (C↓), 123.93 (CH ↑), 123.11 (C↓), 120.42 (C↓), 119.87 (C ↓),
118.69 (CH ↑), 116.72 (CN ↓), 108.40 (CH ↑), 107.90 (CH ↑), 55.58 (C ↓), 40.19 (CH ↑)
MS, m/z (Ir/%): 393 (17.18) (M++ 2), 391 (17.34) (M+) with a base peak at 237 (100)
IR, ν˜/cm−1: 3462, 3399, 3324, 3273, 3189 (2 NH2), 3051, 3032 (CH), 2184 (CN)
1H NMR (DMSO-d6), δ: 7.74–6.70 (m, 9H, 3Ph), 7.06 (bs, 2H, NH2-2, D2O exchangeable ), 5.78 (bs, 2H, NH2-7,
D2O exchangeable), 4.79 (s, 1H, pyran ring), 2.25 (s, 3H, CH3)
Ve
13C NMR (DMSO-d6), δ: 160.12 (C), 144.79 (C), 142.85 (C), 142.71 (C), 135.81 (C), 129.07 (CH), 127.40 (CH),
127.33 (CH), 123.92 (C), 123.30 (CH), 121.90 (C), 120.57 (C), 118.46 (CH), 117.45 (CN), 108.25 (CH), 107.88 (CH),
56.15 (C), 40.41 (CH), 20.49 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 129.07 (CH ↑), 127.40 (CH ↑), 127.33 (CH ↑), 123.30 (CH
↑), 118.46 (CH ↑), 108.25 (CH ↑), 107.88 (CH ↑), 40.41 (CH ↑), 20.49 (CH3 ↑)
13C NMR-DEPT spectrum at 90◦ only CH signals are (up), δ: 129.07 (CH ↑), 127.40 (CH ↑), 127.33 (CH ↑), 123.30
(CH ↑), 118.46 (CH ↑), 108.25 (CH ↑), 107.88 (CH ↑), 40.41 (CH ↑)
13C NMR-DEPT spectrum at 45◦ CH, CH2 and CH3 signals are (up), δ: 129.07 (CH ↑), 127.40 (CH ↑), 127.33 (CH
↑), 123.30 (CH ↑), 118.46 (CH ↑), 108.25 (CH ↑), 107.88 (CH ↑), 40.41 (CH ↑)
13CNMR-APT spectrum CH, CH3 (up), CH2, Cq (down), δ: 160.12 (C), 144.79 (C), 142.85 (C ↓), 142.71 (C ↓),
135.81 (C ↓), 129.07 (CH ↑), 127.40 (CH ↑), 127.33 (CH ↑), 123.92 (C ↓), 123.30 (CH ↑), 121.90 (C ↓), 120.57 (C
↓), 118.46 (CH ↑), 117.45 (CN ↓), 108.25 (CH ↑), 107.88 (CH ↑), 56.15 (C ↓), 40.41 (CH ↑), 20.49 (CH3 ↑)
MS, m/z (Ir/%): 327 (28.55) (M+) with a base peak at 237 (100)

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Table 4. (continued)
Compound
Spectral data
Vf
IR, ν˜/cm−1: 3422, 3382, 3330, 3273, 3209 (2 NH2), 3054, 3031 (CH), 2186 (CN)
1H NMR (DMSO-d6), δ: 7.74–6.70 (m, 9H, 3Ph), 7.02 (bs, 2H, NH2-2, D2O exchangeable), 5.76 (bs, 2H, NH2-7,
D2O exchangeable), 4.78 (s, 1H, pyran ring), 3.71 (s, 3H, CH3)
13C NMR (DMSO-d6), δ: 160.13 (C), 158.08 (C), 144.87 (C), 142.73 (C), 137.99 (C), 128.62 (CH), 127.39 (CH),
124.01 (C), 123.40 (CH), 121.95 (C), 120.67 (C), 118.53 (CH), 117.69 (CN), 113.97 (CH), 108.31 (CH), 107.95 (CH),
56.41 (C), 55.01 (CH3), 40.06 (CH)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.62 (CH ↑), 127.39 (CH↑), 123.40 (CH ↑), 118.53 (CH
↑), 113.97 (CH ↑), 108.31 (CH ↑), 107.95 (CH ↑), 55.01 (CH3 ↑), 40.06 (CH ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.62 (CH ↑), 127.39 (CH↑), 123.40 (CH ↑), 118.53 (CH ↑),
113.97 (CH ↑), 108.31 (CH ↑), 107.95 (CH ↑), 40.06 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.62 (CH ↑), 127.39 (CH ↑), 123.40 (CH ↑), 118.53
(CH ↑), 113.97 (CH ↑), 108.31 (CH ↑), 107.95 (CH ↑), 55.01 (CH3 ↑), 40.06 (CH ↑)
13CNMR-APT spectrum CH, CH3 (up), CH2, Cq (down), δ: 160.13 (C ↓), 158.08 (C ↓), 144.87 (C↓), 142.73 (C ↓),
137.99 (C ↓), 128.62 (CH ↑), 127.39 (CH ↑), 124.01 (C↓), 123.40 (CH ↑), 121.95 (C↓), 120.67 (C↓), 118.53 (CH ↑),
117.69 (CN ↓), 113.97 (CH↑), 108.31 (CH ↑), 107.95 (CH ↑), 56.41 (C ↓), 55.01 (CH3 ↑), 40.06 (CH ↑)
IR, ν˜/cm−1: 3460, 3360, 3310, 3273, 3198 (2 NH2), 3074, 3021 (CH), 2188 (CN)
Vg
Vh
1H NMR (DMSO-d6), δ: 8.22–6.74 (m, 9H, 3Ph), 7.27 (bs, 2H, NH2-2, D2O exchangeable), 5.82 (bs, 2H, NH2-7,
D2O exchangeable), 5.11 (s, 1H, pyran ring)
13C NMR (DMSO-d6), δ: 160.49 (C), 153.16 (C), 146.42 (C), 144.95 (C), 143.05 (C), 129.90 (CH), 128.80 (CH),
127.64 (C), 124.02 (CH), 123.02 (CH), 122.23 (C), 120.29 (C), 118.97 (CH), 116.09 (CN), 108.60 (CH), 107.98 (CH),
55.05 (C), 40.07 (CH)
MS, m/z (Ir/%): 358 (3.19) (M+) with a base peak at 55 (100)
IR, ν˜/cm−1: 3464, 3361, 3315, 3273, 3201 (2 NH2), 3074, 3021, 2877, 2861, 2833 (CH), 2188 (CN)
1H NMR (DMSO-d6), δ: 7.73–6.70 (m, 9H, 3Ph), 7.02 (bs, 2H, NH2-7, D2O exchangeable), 5.77 (bs, 2H, NH2-2,
D2O exchangeable), 4.73 (s, 1H, pyran ring), 3.71–3.69 (m, 4H, 2CH2), 3.05–3.04 (m, 4H, 2CH2)
13C NMR (DMSO-d6), δ: 160.03 (C), 149.78 (C), 144.77 (C), 142.62 (C7), 136.55 (C), 128.21 (CH), 128.03 (CH),
127.23 (C), 123.47 (CH), 121.83 (C), 120.67 (C), 118.38 (CH), 117.75 (CN), 115.13 (CH), 108.18 (CH), 107.86 (CH),
65.98 (CH2), 56.37 (C), 48.35 (CH2), 39.97 (CH)
MS, m/z (Ir/%): 398 (96.32) (M+) with a base peak at 236 (100)
Table 5. Characterisation data of compounds VII
Compound
Formula
Mr
Colour
Yield/%
M.p./◦C
VIIa
VIIb
VIIc
VIId
VIIe
VIIf
VIIg
C
22H19NO3
345.39
363.38
379.84
424.29
359.42
375.42
390.39
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
78
81
83
80
79
80
77
150
155
160
162
152
161
166
C22H18FNO3
C22H18ClNO3
C22H18BrNO3
C23H21NO3
C23H21NO4
C22H18N2O5
Fig. 4. Synthesis of ethyl 2,7-diamino-4H-benzo[h]chromene-3-carboxylate derivatives (VIIIa–VIIIg).
diamino-4H-benzo[h]chromene-3-carboxylate (VIIIa–
VIIIg) derivatives shown in Fig. 4. The composition,
properties and spectral data of the corresponding
products VIII are summarised in Tables 7 and 8.
Structures VII and VIII were established on the
basis of IR, 1H NMR, 13C NMR, 13C NMR-DEPT,
13C NMR-APT and MS data. The IR spectra of VIIa–
VIIg and VIIIa–VIIIg showed the appearance of an

A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
1285
NH2 stretch at 3469–3380 cm−1, 3319–3270 cm−1
CO stretch at 1687–1664 cm−1 for VIIa–VIIg and
a
13C NMR spectra of VIIa–VIIg and VIIIa–VIIIg re-
vealed the presence of 4H signals at δ of 5.19–4.88 (s,
1H, H-4) and 40.15–39.12 (C-4). In compounds VIIa–
VIIg and VIIIa–VIIIg the ester group gave H signals
at δ of 4.05–3.98 (q, 2H, CH2, J = 7.0–7.2 Hz), 1.12–
NH2 stretch at 3470–3417 cm−1, 3390–3362 cm−1
,
3358–3324 cm−1, 3291–3250 cm−1 and a CO stretch
1
at 1679–1667 cm−1 for VIIIa–VIIIg. The 1H and
Table 6. Spectral data of compounds VII
Compound
Spectral data
VIIa
IR, ν˜/cm−1: 3385, 3270 (NH2), 3067, 3029, 2982, 2900, 2876 (CH), 1664 (CO)
1H NMR (DMSO-d6), δ: 8.31–7.10 (m, 11H, 3Ph), 7.79 (bs, 2H, NH2, D2O exchangeable), 5.03 (s, 1H, pyran ring),
4.00 (q, 2H, J = 7.1 Hz, CH2), 1.11 (t, 3H, J = 7.1 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.14 (CO), 160.70 (C), 147.73 (C), 142.74 (C), 132.41 (C), 128.11 (CH), 127.55 (CH),
127.23 (CH), 126.50 (CH), 126.39 (CH), 126.36 (CH), 125.92 (CH), 123.62 (CH), 122.73 (C), 120.87 (C), 120.60
(CH), 76.31 (C), 58.52 (CH2), 39.99 (CH), 14.18 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.11 (CH ↑), 127.55 (CH ↑), 127.23 (CH ↑), 126.50 (CH
↑), 126.39 (CH ↑), 126.36 (CH ↑), 125.92 (CH ↑), 123.62 (CH ↑), 120.60 (CH ↑), 58.52 (CH2 ↓), 39.99 (CH ↑), 14.18
(CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.11 (CH ↑), 127.55 (CH ↑), 127.23 (CH ↑), 126.50 (CH ↑),
126.39 (CH ↑), 126.36 (CH ↑), 125.92 (CH ↑), 123.62 (CH ↑), 120.60 (CH ↑), 39.99 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.11 (CH ↑), 127.55 (CH ↑), 127.23 (CH ↑), 126.50
(CH ↑), 126.39 (CH ↑), 126.36 (CH ↑), 125.92 (CH ↑), 123.62 (CH ↑), 120.60 (CH ↑), 58.52 (CH2 ↑), 39.99 (CH ↑),
14.18 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.14 (CO ↓), 160.70 (C ↓), 147.73 (C ↓), 142.74 (C↓), 132.41
(C↓),128.11 (CH ↑), 127.55 (CH ↑), 127.23 (CH ↑), 126.50 (CH ↑), 126.39 (CH ↑), 126.36 (CH ↑), 125.92 (CH ↑),
123.62 (CH ↑), 122.73 (C↓), 120.87 (C↓), 120.60 (CH ↑), 76.31 (C ↓), 58.52 (CH2 ↓), 39.99 (CH ↑), 14.18 (CH3 ↑)
MS, m/z (Ir/%): 345 (23.84) (M+) with a base peak at 237 (100)
VIIb
IR, ν˜/cm−1: 3385, 3288 (NH2), 3077, 3063, 2987, 2978, 2960, (CH), 1668 (CO)
1H NMR (DMSO-d6), δ: 8.33–7.03 (m, 10H, 3Ph), 7.82 (bs, 2H, NH2, D2O exchangeable), 5.06 (s, 1H, pyran ring),
4.01 (q, 2H, J = 7.2 Hz, CH2), 1.10 (t, 3H, J = 7.2 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.10 (CO), 161.46 (C), 160.66 (C), 144.00 (C), 142.72 (C), 132.47 (C), 129.06 (CH),
129.00 (CH), 128.24 (CH), 127.58 (CH), 126.45 (CH), 123.73 (CH), 122.75 (C), 120.64 (C), 120.62 (CH), 114.88
(CH), 114.71 (CH), 76.25 (C), 58.58 (CH2), 40.02 (CH), 14.21 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 129.06 (CH ↑), 129.00 (CH ↑), 128.24 (CH ↑), 127.58 (CH
↑), 126.45 (CH ↑), 123.73 (CH ↑), 120.62 (CH ↑), 114.88 (CH ↑), 114.75 (CH ↑), 58.58 (CH2 ↓), 40.02 (CH ↑), 14.21
(CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 129.06 (CH ↑), 129.00 (CH ↑), 128.24 (CH ↑), 127.58 (CH ↑),
126.45 (CH ↑), 123.73 (CH ↑), 120.62 (CH ↑), 114.88 (CH ↑), 114.75 (CH ↑), 40.02 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 129.06 (CH ↑), 129.00 (CH ↑), 128.24 (CH ↑), 127.58
(CH ↑), 126.45 (CH ↑), 123.73 (CH ↑), 120.62 (CH ↑), 114.88 (CH ↑), 114.75 (CH ↑), 58.58 (CH2 ↑), 40.02 (CH ↑),
14.21 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.10 (CO ↓), 161.46 (C ↓), 160.66 (C ↓), 144.00 (C ↓), 142.72
(C ↓), 132.47 (C ↓), 129.06 (CH ↑), 129.00 (CH ↑), 128.24 (CH ↑), 127.58 (CH ↑), 126.45 (CH ↑), 123.73 (CH ↑),
122.75 (C ↓), 120.64 (C ↓), 120.62 (CH ↑), 114.88 (CH ↑), 114.75 (CH ↑), 76.25 (C ↓), 58.58 (CH2 ↓), 40.02 (CH ↑),
14.21 (CH3 ↑)
MS, m/z (Ir/%): 363 (1.64) (M+) with a base peak at 68 (100)
VIIc
IR, ν˜/cm−1: 3469, 3318 (NH2), 3079, 3058, 2977, 2955, 2903 (CH), 1678 (CO)
1H NMR (DMSO-d6), δ: 8.32–7.25 (m, 10H, 3Ph), 7.83 (bs, 2H, NH2), 5.06 (s, 1H, pyran ring), 4.01 (q, 2H, J =
7.1 Hz, CH2), 1.10 (t, 3H, J = 7.1 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.01 (CO), 160.66 (C), 146.75 (C), 142.73 (C), 132.50 (C), 130.46 (C), 129.12 (CH),
128.08 (CH), 127.57 (CH), 126.48 (CH), 126.46 (CH), 126.37 (CH), 123.74 (CH), 122.72 (C), 120.63 (C), 120.26
(CH), 75.92 (C), 58.60 (CH2), 39.44 (CH), 14.20 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 129.12 (CH ↑), 128.08 (CH ↑), 127.57 (CH ↑), 126.48 (CH
↑), 126.46 (CH ↑), 126.37 (CH ↑), 123.74 (CH ↑), 120.26 (CH ↑), 58.60 (CH2 ↓), 39.44 (CH ↑), 14.20 (CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 129.12 (CH ↑), 128.08 (CH ↑), 127.57 (CH ↑), 126.48 (CH ↑),
126.46 (CH ↑), 126.37 (CH ↑), 123.74 (CH ↑), 120.26 (CH ↑), 39.44 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 129.12 (CH ↑), 128.08 (CH ↑), 127.57 (CH ↑), 126.48
(CH ↑), 126.46 (CH ↑), 126.37 (CH ↑), 123.74 (CH ↑), 120.26 (CH ↑), 58.60 (CH2 ↑), 39.44 (CH ↑), 14.20 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.01 (CO ↓), 160.66 (C ↓), 146.75 (C ↓), 142.73 (C ↓), 132.50
(C ↓), 130.46 (C ↓), 129.12 (CH ↑), 128.08 (CH ↑), 127.57 (CH ↑), 126.48 (CH ↑), 126.46 (CH ↑), 126.37 (CH ↑),
123.74 (CH ↑), 122.72 (C ↓), 120.63 (C ↓), 120.26 (CH ↑), 75.92 (C ↓), 58.60 (CH2 ↓), 39.44 (CH ↑), 14.20 (CH3 ↑)
MS, m/z (Ir/%): 381(8.18)(M++ 2), 379 (24.72) (M+) with a base peak at 269 (100)

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Table 6. (continued)
Compound
Spectral data
VIId
IR, ν˜/cm−1: 3463, 3319 (NH2), 3077, 3069, 2978, 2976, 2931 (CH), 1677 (CO)
1H NMR (DMSO-d6), δ: 8.33–7.20 (m, 10H, 3Ph), 7.84 (bs, 2H, NH2), 5.05 (s, 1H, pyran ring), 4.01(q, 2H, J = 7.2
Hz, CH2), 1.10 (t, 3H, J = 7.2 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.02 (CO), 160.87 (C), 147.18 (C), 142.74 (C), 132.51 (C), 131.12 (CH), 129.54 (CH),
127.58 (CH), 126.49 (CH), 126.46 (CH), 123.75 (CH), 122.73 (CH), 120.64 (C), 120.38 (C), 120.19 (CH), 119.03
(C), 75.86 (C), 58.61 (CH2), 39.29 (CH), 14.22 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 131.12 (CH ↑), 129.54, (CH ↑), 127.58 (CH ↑), 126.49 (CH
↑), 126.46 (CH ↑), 123.75 (CH ↑), 122.73 (CH ↑), 120.19 (CH ↑), 58.61 (CH2 ↓), 39.29 (CH ↑), 14.22 (CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 131.12 (CH ↑), 129.54, (CH ↑), 127.58 (CH ↑), 126.49 (CH ↑),
126.46 (CH ↑), 123.75 (CH ↑), 122.73 (CH ↑), 120.19 (CH ↑), 39.29 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 131.12 (CH ↑), 129.54, (CH ↑), 127.58 (CH ↑),
126.49 (CH ↑), 126.46 (CH ↑), 123.19 (CH ↑), 122.73 (CH ↑), 120.19 (CH ↑), 39.29 (CH ↑), 58.61 (CH2 ↑), 39.29
(CH ↑), 14.22 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.02 (CO ↓), 160.87 (C ↓), 147.18 (C ↓), 142.74 (C ↓), 132.51
(C ↓), 131.12 (CH ↑), 129.54, (CH ↑), 127.58 (CH ↑), 126.49 (CH ↑), 126.46 (CH ↑), 123.75 (CH ↑), 122.73 (CH ↑),
120.64 (C ↓), 120.38 ( C ↓), 120.19 ( CH ↑), 119.03 (C ↓), 75.86 (C ↓), 58.61 (CH2 ↓), 39.29 (CH ↑), 14.22 (CH3 ↑)
MS, m/z (Ir/%): 425 (12.18) (M++ 2), 423 (12.83) (M+) with a base peak at 269 (100)
IR, ν˜/cm−1: 3380, 3287 (NH2), 3079, 3063, 2996, 2977, 2901 (CH), 1681 (CO)
VIIe
1H NMR (DMSO-d6), δ: 8.30–7.02 (m, 10H, 3Ph), 7.76 (bs, 2H, NH2, D2O exchangeable), 4.98 (s, 1H, pyran ring),
3.99 (q, 2H, J = 7.2 Hz, CH2), 2.19 (s, 3H, CH3), 1.11 (t, 3H, J = 7.2 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.19 (CO), 160.65 (C), 144.80 (C), 142.70 (C), 134.87 (C), 132.38 (C), 128.66 (CH),
127.54 (CH), 127.11 (CH), 126.52 (CH), 126.35 (CH), 126.31 (CH), 123.56 (CH), 122.74 (C), 121.09 (C), 120.58
(CH), 76.42 (C), 58.53 (CH2), 39.44 (CH), 20.44 (CH3), 14.22 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.66 (CH ↑), 127.54 (CH ↑), 127.11 (CH ↑), 126.52 (CH
↑), 126.35 (CH ↑), 126.31 (CH ↑), 123.56 (CH ↑), 120.58 (CH ↑), 58.53 (CH2 ↓), 39.44 (CH ↑), 20.44 (CH3 ↑), 14.22
(CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.66 (CH ↑), 127.54 (CH ↑), 127.11 (CH ↑), 126.52 (CH ↑),
126.35 (CH ↑), 126.31 (CH ↑), 123.56 (CH ↑), 120.58 (CH ↑), 39.44 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.66 (CH ↑), 127.54 (CH ↑), 127.11 (CH ↑), 126.52
(CH ↑), 126.35 (CH ↑), 126.31 (CH ↑), 123.56 (CH ↑), 120.58 (CH ↑), 58.53 (CH2 ↑), 39.44 (CH ↑), 20.44 (CH3 ↑),
14.22 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.19 (CO↓), 160.65 (C↓), 144.80 (C ↓),142.70 (C ↓),134.87 (C
↓), 132.38 (C↓), 128.66 (CH ↑), 127.54 (CH ↑), 127.11 (CH ↑), 126.52 (CH ↑), 126.35 (CH ↑), 126.31 (CH ↑), 123.56
(CH ↑), 122.74 (C ↓), 121.09 (C ↓), 120.58 (CH ↑), 76.42 (C ↓), 58.53 (CH2 ↓), 39.44 (CH ↑), 20.44 (CH3 ↑), 14.22
(CH3 ↑)
MS, m/z (Ir/%): 359 (15.84) (M+) with a base peak at 273 (100)
VIIf
IR, ν˜/cm−1: 3467, 3325 (NH2), 3081, 3062, 2993, 2983, 2904 (CH), 1687 (CO)
1H NMR (DMSO-d6), δ: 8.31–6.77 (m, 10H, 3Ph), 7.76 (bs, 2H, NH2), 4.98 (s, 1H, pyran ring), 4.00 (q, 2H, J =
7.2 Hz, CH2), 3.66 (s, 3H, OCH3), 1.12 (t, 3H, J = 7.2 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.28 (CO), 160.67 (C), 157.48 (C), 142.73 (C), 140.00 (C), 132.42 (C), 128.25 (CH),
127.60 (CH), 126.62 (CH), 126.42 (CH), 126.35 (CH), 123.63 (CH), 122.81 (C), 121.29 (C), 120.64 (CH), 113.54
(CH), 76.66 (C), 58.59 (CH2), 54.90 (CH3), 39.17 (CH), 14.30 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.25 (CH ↑), 127.60 (CH ↑), 126.62 (CH ↑), 126.42 (CH
↑), 126.35 (CH ↑), 123.63 (CH ↑), 120.64 (CH ↑), 113.54 (CH ↑), 58.59 (CH2 ↓), 54.90 (CH3 ↑), 39.17 (CH ↑), 14.30
(CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.25 (CH ↑), 127.60 (CH ↑), 126.62 (CH ↑), 126.42 (CH ↑),
126.35 (CH ↑), 123.63 (CH ↑), 120.64 (CH ↑), 113.54 (CH ↑), 39.17 (CH ↑)
13C NMR-DEPT spectrum at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.25 (CH ↑), 127.60 (CH ↑), 126.62
(CH ↑), 126.42 (CH ↑), 126.35 (CH ↑), 123.63 (CH ↑), 120.64 (CH ↑), 113.54 (CH ↑), 58.59 (CH2 ↑), 54.90 (CH3 ↑),
39.17 (CH ↑), 14.30 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.28 (CO ↓), 160.67 (C ↓), 157.48 (C ↓), 142.73 (C↓), 140.00
(C ↓), 132.42 (C↓), 128.25 (CH ↑), 127.60 (CH ↑), 126.62 (CH ↑), 126.42 (CH ↑), 126.35 (CH ↑), 123.63 (CH ↑),
122.81 (C ↓), 121.29 (C ↓), 120.64 (C ↑), 113.54 (CH ↑), 76.66 (C ↓), 58.59 (CH2 ↓), 54.90 (CH3 ↑), 39.17 (CH ↑),
14.30 (CH3 ↑)
MS, m/z (Ir/%): 375 (11.32) (M+) with a base peak at 269 (100)
VIIg
IR, ν˜/cm−1: 3467, 3317 (NH2), 3085, 3066, 2995, 2977, 2908 (CH), 1687 (CO)
1H NMR (DMSO-d6), δ: 8.47–7.13 (m, 10H, 3Ph), 8.00 (bs, 2H, NH2, D2O exchangeable), 5.28 (s, 1H, pyran ring),
3.99 (q, 2H, J = 7 Hz, CH2), 1.11 (t, 3H, J = 7 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.75 (CO), 160.67 (C), 154.86 (C), 146.00 (C), 142.30 (C), 132.42 (C), 128.74 (CH),
127.66 (CH), 126.66 (CH), 126.42 (CH), 126.37 (CH), 125.13 (CH),123.65 (CH), 122.82 (C), 121.27 (C), 120.61
(CH), 76.65 (C), 58.58 (CH2), 40.03 (CH), 14.30 (CH3)
MS, m/z (Ir/%): 390 (13.46) (M+) with a base peak at 269 (100)

A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
Table 7. Characterisation data of compounds VIII
1287
Compound
Formula
Mr
Colour
Yield/%
M.p./◦C
VIIIa
VIIIb
VIIIc
VIIId
VIIIe
VIIIf
VIIIg
C
22H20N2O3
360.41
378.40
394.85
439.30
374.43
390.43
405.40
Grey
Grey
Red
Red
Red
Red
Red
74
77
73
75
76
75
71
203, 203a
210
C22H19FN2O3
C22H19ClN2O3
C22H19BrN2O3
C23H22N2O3
C23H22N2O4
C22H19N3O5
185, 185a
188
196
192, 192a
213
a) According to Abd-El-Aziz et al. (2004).
Fig. 5. Anti-tumour activity of 4H-benzo[h]chromene derivatives: MCF-7 (blue); HCT-116 (red) and HepG-2 (green).
1.06 (t, 3H, CH3, J = 7.0–7.2 Hz) with the correspond-
ing signals in the 13C spectra at δ of 58.65–58.46 (CH2)
and 14.32–14.18 (CH3), respectively. The 13C NMR-
DEPT spectra at 45,◦ 90,◦ 135,◦ 13C NMR-APT spec-
tra and the mass spectra of compounds VII and VIII
provided additional evidence in support of the pro-
posed structures. In addition, the 1H NMR spectra
for compounds III and V showed that the NH2 pro-
tons resonated at δ of 7.37–7.02 (sharp singlet), while
compounds VII and VIII showed that the NH2 pro-
tons resonated at δ of 8.00–7.71 (broad singlet lower
field), respectively. This de-shielding is a result of the
toxicity evaluation using viability assay was performed
at the Regional Centre for Mycology & Biotechnology
(RCMP), Al-Azhar University (Cairo, Egypt), using
vinblastine and colchicine as standard drugs under
different concentrations (50 μg mL−1, 25 μg mL−1
,
12.5 μg mL−1, 6.25 μg mL−1, 3.125 μg mL−1, 1.56
μg mL−1 and 0 μg mL−1). The inhibitory activity of
the synthetic compounds IIIa–IIIh, Va–Vh, VIIa–VIIg
and VIIIa–VIIIg against the three cell lines MCF-7,
HCT-116 and HepG-2 is given in Table 9 and Fig. 5.
SAR studies
—
replacement of the CN group in III and V by the C
O
—
—
—
group in VII and VIII whose C O anisotropy would
The preliminary SAR study focused on the ef-
fect of the substituent at the phenyl group at the
4-position and the substituent at the 3- and 7-
positions of the 4H-benzo[h]chromene moiety, on the
anti-tumour activities of the synthesised compounds.
In a comparison of the cytotoxic activities of the
four series (IIIa–IIIh, Va–Vh, VIIa–VIIg and VIIIa–
VIIIg) against breast adenocarcinoma (MCF-7), it
was found that, for the first series IIIa–IIIh and the
second series Va–Vh, the highest growth inhibitory
effect was associated with the unsubstituted phenyl
IIIa and 4-methoxyphenyl Vf analogues with IC50
of 17.5 μg mL−1 and 14.8 μg mL−1, respectively,
which exhibited good activity relative to colchicine
(IC50 = 17.7 μg mL−1) and more reduction in po-
tency with the other derivatives IIIg, IIIe, IIIb, IIIc,
de-shield these protons in addition to the protons in-
—
volved in the hydrogen bonding with the C O group.
—
This was also, supported by the X-ray single crystal
data (Al-Dies et al., 2012, El-Agrody et al., 2012).
Anti-tumour assays
Compounds IIIa–IIIh, Va–Vh, VIIa–VIIg and
VIIIa–VIIIg were evaluated for human tumour cell
growth inhibitory activity against three cell lines:
breast adenocarcinoma (MCF-7), lung carcinoma
(HCT-116) and hepatocellular carcinoma (HepG-2).
The measurements of cell growth and the viabilities
were determined as described in the literature (Moss-
man, 1983; Rahman et al., 2001). The in-vitro cyto-

1288
A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
IIId, IIIf, IIIh and Vg, Ve, Va, Vb, Vc, Vd, Vh with
IC50 = 41.19–47.5 μg mL−1 or > 50 μg mL−1 and
18.4–43.7 μg mL−1 or > 50 μg mL−1, respectively,
(electron-donating) at the para-position on the phenyl
ring at the 4-position of the 4H-benzo[h]chromene
moiety with NH2-7 (electron-donating) of the sec-
ond series is preferred for anti-tumour activity than
the other substituent. Replacement of the electron-
withdrawing cyano group of the first series III and
the second series V by the ester group at the
as compared with vinblastine (IC50 = 6.1 μg mL−1
)
and colchicine (IC50 = 17.7 μg mL−1), suggest-
ing that the unsubstituted phenyl (electron-donating)
with H-7 of the first series and the methoxy group
Table 8. Spectral data of compounds VIII
Compound
Spectral data
VIIIa
IR, ν˜/cm−1: 3436, 3390, 3350, 3275 (2 NH2), 3064, 3021, 2983 (CH), 1677 (CO)
1H NMR (DMSO-d6), δ: 7.78–6.69 (m, 10H, 3Ph), 7.76 (bs, 2H, NH2-2, D2O exchangeable), 5.76 (bs, 2H, NH2-7,
D2O exchangeable), 4.98 (s, 1H, pyran ring), 3.99 (q, 2H, J = 7 Hz, CH2), 1.08 (t, 3H, J = 7 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.22 (CO), 160.91 (C), 147.89 (C), 144.77 (C), 142.89 (C), 128.05 (CH), 127.19 (CH
↑), 127.17 (CH), 125.83 (CH), 124.00 (C), 123.67 (CH), 121.81 (C), 120.39 (C), 118.38 (CH), 108.01 (CH), 107.91
(CH), 76.27 (C), 58.47 (CH2), 40.00 (CH), 14.19 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.05 (CH ↑), 127.19 (CH ↑), 127.17 (CH ↑), 125.83 (CH
↑), 123.67 (CH ↑), 118.38 (CH ↑), 108.01 (CH ↑), 107.91 (CH ↑), 58.47 (CH2 ↓), 40.00 (CH ↑), 14.19 (CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.05 (CH ↑), 127.19 (CH ↑), 127.17 (CH ↑), 125.83 (CH ↑),
123.67 (CH ↑), 118.38 (CH ↑), 108.01 (CH ↑), 107.91 (CH ↑), 40.00 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.05 (CH ↑), 127.19 (CH ↑), 127.17 (CH ↑), 125.83
(CH ↑), 123.67 (CH ↑), 118.38 (CH ↑), 108.01 (CH ↑), 107.91 (CH ↑), 58.47 (CH2 ↑), 40.00 (CH ↑), 14.19 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.22 (CO ↓), 160.91 (C ↓), 147.89 (C ↓), 144.77 (C ↓), 142.89
(C ↓), 128.05 (CH ↑), 127.19 (CH ↑), 127.17 (CH ↑), 125.83 (CH ↑), 124.00 (C ↓), 123.67 (CH ↑), 121.81 (C ↓),
120.39 (C ↓), 118.38 (CH ↑), 108.01 (CH ↑), 107.91 (CH ↑), 76.27 (C ↓), 58.47 (CH2 ↓), 40.00 (CH ↑), 14.19 (CH3 ↑)
IR, ν˜/cm−1: 3440, 3390, 3354, 3222 (2 NH2), 3060, 2988, 2932 (CH), 1679 (CO)
VIIIb
VIIIc
VIIId
1H NMR (DMSO-d6), δ: 7.78–6.78 (m, 9H, 3Ph), 7.82 (bs, 2H, NH2-2), 5.79 (bs, 2H, NH2-7), 5.18 (s, 1H, pyran
ring), 3.99 (q, 2H, J = 7 Hz, CH2), 1.10 (t, 3H, J = 7 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.24 (CO), 160.93 (C), 160.14 (C), 146.93 (C), 144.01 (C), 142.89 (C), 129.08 (CH),
127.28 (CH), 124.03 (C), 123.68 (CH), 121.90 (C), 120.39 (C), 118.39 (CH), 114.89 (CH), 108.15 (CH), 107.95 (CH),
76.28 (C), 58.56 (CH2), 40.07 (CH), 14.19 (CH3)
MS, m/z (Ir/%): 378 (13.46) (M+) with a base peak at 264 (100)
IR, ν˜/cm−1: 3417, 3390, 3324, 3250 (2 NH2), 3065, 2982, 2931 (CH), 1667 (CO)
1H NMR (DMSO-d6), δ: 7.77–6.72 (m, 9H, 3Ph), 7.81 (bs, 2H, NH2-2, D2O exchangeable), 5.79 (bs, 2H, NH2-7,
D2O exchangeable), 5.02 (s, 1H, pyran ring), 4.00 (q, 2H, J = 7.2 Hz, CH2), 1.10 (t, 3H, J = 7.2 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.10 (CO), 160.88 (C), 146.93 (C), 144.82 (C), 142.90 (C), 130.38 (C), 129.08 (CH),
128.23 (CH), 127.28 (CH), 124.01 (C), 123.56 (CH), 121.90 (C), 119.79 (C), 118.53 (CH), 108.15 (CH), 107.95 (CH),
75.90 (C), 58.56 (CH2), 40.01 (CH), 14.22 (CH3)
MS, m/z (Ir/%): 398 (1.11)(M++ 2), 396 (3.46) (M+) with a base peak at 264 (100)
IR, ν˜/cm−1: 3449, 3377, 3355, 3271 (2 NH2), 3062, 2963 (CH), 1679 (CO)
1H NMR (DMSO-d6), δ: 7.76–6.71 (m, 9H, 3PH), 7.15 (bs, 2H, NH2-2, D2O exchangeable), 5.80 (bs, 2H, NH2-7,
D2O exchangeable), 4.88 (s, 1H, pyran ring), 3.99 (q, 2H, J = 7 Hz, CH2), 1.06 (t, 3H, J = 7 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.21 (CO), 160.21 (C), 145.16 (C), 144.83 (C), 142.79 (C), 131.45 (CH), 129.76 (CH),
127.33 (CH), 123.91 (C), 123.08 (CH), 121.98 (C), 120.39 (C), 119.84 (C), 118.68 (CH), 108.35 (CH), 107.84 (CH),
76.25 (C), 58.46 (CH2), 40.15 (CH), 14.19 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 131.45 (CH ↑), 129.76 (CH ↑), 127.43 (CH ↑), 123.08 (CH
↑), 118.68 (CH ↑), 108.35 (CH ↑), 107.84 (CH ↑), 58.46 (CH2 ↓), 40.15 (CH ↑), 14.19 (CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 131.45 (CH ↑), 129.76 (CH ↑), 127.43 (CH ↑), 123.08 (CH ↑),
118.68 (CH ↑), 108.35 (CH ↑), 107.84 (CH ↑), 40.15 (CH ↑)
13C NMR-DEPT at 45◦ CH, CH2 and CH3 signals are (up), δ: 131.45 (CH ↑), 129.76 (CH ↑), 127.43 (CH ↑), 123.08
(CH ↑), 118.68 (CH ↑), 108.35 (CH ↑), 107.84 (CH ↑), 58.46 (CH2 ↑), 40.15 (CH ↑), 14.19 (CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.21 (CO ↓), 160.21 (C ↓), 145.16 (C ↓), 144.83 (C ↓), 142.79
(C ↓), 131.45 (CH ↑), 129.76 (CH ↑), 127.43 (CH ↑), 123.91 (C ↓), 123.08 (CH ↑), 121.98 (C ↓), 120.39 (C ↓), 119.84
(C ↓), 118.68 (CH ↑), 108.35 (CH ↑), 107.84 (CH ↑), 76.25 (C ↓), 58.46 (CH2 ↓), 40.15 (CH ↑), 14.19 (CH3 ↑)
MS, m/z (Ir/%): 440 (1.11) (M++ 2), 438 (1.16) (M+) with a base peak at 236 (100)
VIIIe
IR, ν˜/cm−1: 3439, 3362, 3358, 3272 (2 NH2), 3061, 2932, 2859 (CH), 1678 (CO)
1H NMR (DMSO-d6), δ: 7.72–6.68 (m, 9H, 3Ph), 7.71 (bs, 2H, NH2-2), 5.75 (bs, 2H, NH2-7), 4.93 (s, 1H, pyran
ring), 3.98 (q, 2H, J = 7 Hz, CH2,), 2.19 (s, 3H, CH3,), 1.08 (t, 3H, J = 7 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.22 (CO), 160.23 (C), 144.80 (C), 144.75 (C), 142.80 (C), 134.75 (C), 128.60 (CH),
127.04 (CH), 126.35 (CH), 123.94 (C), 123.09 (CH), 121.99 (C), 119.88 (C), 118.68 (CH), 107.95 (CH), 107.87 (CH),
76.35 (C), 58.49 (CH2), 40.07 (CH), 20.44 (CH3), 14.32 (CH3)
MS, m/z (Ir/%): 374 (42.35) (M+) with a base peak at 248 (100)

A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
1289
Table 8. (continued)
Compound
Spectral data
VIIIf
IR, ν˜/cm−1: 3463, 3366, 3342, 3257 (2 NH2), 3071, 2023, 2982 (CH), 1667 (CO)
1H NMR (DMSO-d6), δ: 7.73–6.69 (m, 9H, 3Ph), 7.70 (bs, 2H, NH2-2), 5.74 (bs, 2H, NH2-7), 4.93 (s, 1H, pyran
ring), 3.99 (q, 2H, J = 7 Hz, CH2), 3.66 (s, 3H, CH3), 1.10 (t, 3H, J = 7 Hz, CH3)
13C NMR (DMSO-d6), δ: 168.35 (CO), 160.88 (C), 157.42 (C), 144.83 (C), 142.91 (C), 140.17 (C), 128.18 (CH),
127.21 (CH), 124.07 (C), 123.80 (CH), 121.82 (C), 120.80 (C), 118.39 (CH), 113.49 (CH), 108.04 (CH), 107.98 (CH),
76.63 (C), 58.54 (CH2), 54.90 (CH3), 39.12 (CH), 14.32 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.18 (C ↑), 127.21 (CH ↑), 123.80 (CH ↑), 118.39 (CH
↑), 113.49 (CH ↑), 108.04 (CH ↑), 107.98 (CH ↑), 58.54 (CH2 ↓), 54.90 (CH3 ↑), 39.12 (CH ↑), 14.32 (CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.18 (C ↑), 127.21 (CH ↑), 123.80 (CH ↑), 118.39 (CH ↑),
113.49 (CH ↑), 108.04 (CH ↑), 107.98 (CH ↑), 39.12 (CH ↑)
13C NMR-DEPT spectrum at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.18 (CH ↑), 127.21 (CH ↑), 123.80 (CH
↑), 118.39 (CH ↑), 113.49 (CH↑), 108.04 (CH ↑), 107.98 (CH ↑), 58.54 (CH2 ↑), 54.90 (CH3 ↑), 39.12 (CH ↑), 14.32
(CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 168.35 (CO ↓), 160.88 (C ↓), 157.42 (C ↓), 144.83 (C ↓), 142.91
(C ↓), 140.17 (C ↓), 128.18 (CH ↑), 127.21 (CH ↑), 124.07 (C ↓), 123.80 (CH ↑), 121.82 (C ↓), 120.80 (C ↓), 118.39
(CH ↑), 113.49 (CH ↑), 108.04 (CH ↑), 107.98 (CH ↑), 76.63 (C ↓), 58.54 (CH2 ↓), 54.90 (CH3 ↑), 39.12 (CH ↑),
14.32 (CH3 ↑)
MS, m/z (Ir/%): 390 (1.16) (M+) with a base peak at 56 (100)
VIIIg
IR, ν˜/cm−1: 3470, 3376, 3351, 3291 (2 NH2), 3071, 2023, 2982 (CH), 1670 (CO)
1H NMR (DMSO-d6), δ: 8.14–6.72 (m, 9H, 3Ph), 7.89 (bs, 2H, NH2, D2O exchangeable), 5.81 (bs, 2H, NH2, D2O
exchangeable), 5.19 (s, 1H, pyran ring), 3.99 (q, 2H, J = 7 Hz, CH2), 1.09 (t, 3H, J = 7 Hz, CH3)
13C NMR (DMSO-d6), δ: 167.90 (CO), 160.90 (C), 155.58 (C), 145.72 (C), 144.86 (C), 142.96 (C), 128.54 (CH),
128.23 (CH), 127.40 (CH), 124.01 (C), 123.51 (CH), 123.40 (CH), 122.04 (C), 118.76 (C), 118.70 (CH), 108.28 (CH),
107.95 (CH), 75.22 (C), 58.65 (CH2), 40.03 (CH), 14.20 (CH3)
13C NMR-DEPT at 135◦ CH, CH3 (up), CH2 (down), δ: 128.54 (CH ↑), 128.23 (CH ↑), 127.40 (CH ↑), 123.51 (CH
↑), 123.40 (CH ↑), 118.70 (CH ↑), 108.28 (CH ↑), 107.95 (CH ↑), 58.65 (CH2 ↓), 40.03 (CH ↑), 14.20 (CH3 ↑)
13C NMR-DEPT at 90◦ only CH signals are (up), δ: 128.54 (CH ↑), 128.23 (CH ↑), 127.40 (CH ↑), 123.51 (CH ↑),
123.40 (CH ↑), 118.70 (CH ↑), 108.28 (CH ↑), 107.95 (CH ↑), 40.03 (CH ↑)
13C NMR-DEPT spectrum at 45◦ CH, CH2 and CH3 signals are (up), δ: 128.54 (CH ↑), 128.23 (CH ↑),127.40 (CH
↑), 123.51 (CH ↑), 123.40 (CH ↑), 118.70 (CH ↑), 108.28 (CH ↑), 107.95 (CH ↑), 58.65 (CH2 ↑), 40.03 (CH ↑), 14.20
(CH3 ↑)
13CNMR-APT CH, CH3 (up), CH2, Cq (down), δ: 167.90 (CO ↓), 160.90 (C ↓), 155.58 (C ↓), 145.72 (C ↓), 144.86
(C↓), 142.96 (C ↓), 128.54 (CH ↑), 128.23 (CH ↑), 127.40 (CH ↑), 124.01 (C ↓), 123.51 (CH ↑), 123.40 (CH ↑),
122.04 (C ↓), 118.76 (C ↓), 118.70 (CH ↑), 108.28 (CH ↑), 107.95 (CH ↑), 75.22 (C ↓), 58.65 (CH2 ↓), 40.03 (CH ↑),
14.20 (CH3 ↑)
MS, m/z (Ir/%): 405 (26.39) (M+) with a base peak at 284 (100)
3−position resulted in a marked improvement in po-
tency for the third series VII and the fourth series VIII
against MCF-7, 4-bromophenylVIId, 4-fluorophenyl
VIIb,
In the case of lung carcinoma (HCT-116), the SAR
investigation for the four series revealed compound
VIIb of the third series (IC50 = 2.9 μg mL−1) to
have potent anti-tumour activity against the HCT-116
closet to vinblastine (IC50 = 2.6 μg mL−1); the other
compounds of the four series exhibited moderate to
lower activities, while compounds IIIa, IIId, IIIg, IIIf,
IIIb, IIIh, IIIc, IIIe of the first series (IC50 = 7.1–
33.2 μg mL−1) and Vg, Vf of the second series (IC50
of 30.8 μg mL−1 and 37.8 μg mL−1, respectively)
had the most potent activity against HCT-116 com-
pared to colchicine (IC50 = 42.8 μg mL−1), suggesting
that the unsubstituted phenyl (electron-donating), the
bromo, nitro, fluoro and chloro (electron-withdrawing)
or methoxy, morpholinophenyl and methyl groups
(electron-donating) of the first series with H-7 and
the nitro (electron-withdrawing) and methoxy groups
(electron-donating) of the second series with NH2-7
at the para-position of phenyl ring at the 4-position
are preferred over the other substituent. Replace-
ment of the electron-withdrawing cyano group of
first series III and the second series V by the es-
4-methylphenyl VIIe (IC50 = 3.7–4.7 μg mL−1) and
4-chlorophenyl VIIIc, 4-nitrophenyl VIIIg, analogues
(IC50 of 3.1 μg mL−1 and 5.3 μg mL−1, respectively)
exhibited good activity against MCF-7 compared with
vinblastine (IC50 = 6.1 μg mL−1) and colchicine
(IC50 = 17.7 μg mL−1), while 4-chlorophenyl VIIc and
4-methylphenyl VIIIe with (IC50 of 6.5 μg mL−1 and
8.8 μg mL−1, respectively) have a significant potent
anti-tumour activity in compareison with colchicine,
suggesting that the bulky substituent and the elec-
tronic nature of the substituent (electron-withdrawing
or electron-donating groups) of the third series with
H-7 or the less bulky substituent (electron-withdraw-
ing) of the fourth series with NH2-7 may be the main
factor affecting the potency of these compounds. In
general, the activities against MCF-7 of the four se-
ries were decreased in the descending order:VII, VIII,
V , III.

1290
A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
Table 9. SAR of 4-aryl group, 3-, 7-positions and IC50 of target compounds against different cell lines in comparison with the
standard as measured with the MTT methoda
Compound
R
MCF-7
HCT-116
HepG-2
Compound
R
MCF-7
22.6 0.1
HCT-116
HepG-2
IIIa
IIIb
IIIc
IIId
IIIe
IIIf
IIIg
IIIh
Va
Vb
Vc
Vd
Ve
H
F
Cl
17.5
0.1
0.0
7.1
21.2
28.8
9.6
33.2
12.8
9.8
0.1
0.3
0.1
0.1
0.0
0.0
0.0
0.0
3.2
35.8
24.3
10.6
32.1
0.0
VIIa
VIIb
VIIc
VIId
VIIe
VIIf
VIIg
VIIIa
VIIIb
VIIIc
VIIId
VIIIe
VIIIf
VIIIg
B
H
F
Cl
8.4
2.9
9.6
5.4
5.6
0.1
0.2
0.7
0.0
0.0
0.0
6.0
5.6
0.3
47.5
42.8
0.1
0.0
0.1
0.1
4.1
6.5
3.7
4.7
48.8
0.1
0.2
0.2
0.0
0.0
0.1
0.4
0.2
0.0
0.1
0.0
0.2
0.0
0.1
0.2
0.1
0.1
0.3
0.1
0.0
W
W
4.1
Br
Br
22.5
2.5
Me
OMe
NO2
A
H
F
Cl
Br
Me
OMe
NO2
A
0.4
0.1
Me
OMe
NO2
H
F
Cl
W
41.9
W
44.9
27.9
35.1
47.3
13.7
10.9
24.3
10.4
8.3
2.8
44.6
20.7
40.7
46.9
44.9
23.6
38.4
22.8
11.1
0.0
0.3
0.1
0.0
0.4
0.1
0.2
0.4
0.1
0.2
W
W
W
W
W
W
W
W
21.3
22.6
22.5
8.5
0.4
0.4
0.0
W
24.6
3.1
40.3
8.8
0.4
0.1
0.1
0.0
45.4
0.0
W
W
W
Br
W
Me
OMe
NO2
–
27.0
39.4
35.2
2.6
0.0
0.1
0.0
0.0
0.1
43.7
14.8
18.4
0.0
0.0
0.1
W
Vf
Vg
Vh
37.8
30.8
0.1
0.2
5.3
6.1
17.7
0.0
0.0
0.0
15.3
4.6
W
W
C
–
42.8
10.6
a) IC50 values expressed in μg mL−1 as mean values of triplicate wells from at least three experiments and are reported as the
mean
standard error; W = weak activity (IC50 > 50 μg mL−1); A = morpholino; B = vinblastine; C = colchicine.
ter group at the 3-position resulted in a marked
increase in potency for the third series VII and
a little reduction in potency for the fourth series
VIII against HCT-116, the 4-fluorophenyl VIIb, 4-
bromophenyl VIId, 4-methylphenyl VIIe, phenyl VIIa,
4-chlorophenyl VIIc analogues for the third series with
para-position of the phenyl ring at the 4-position for
the fourth series VIII, suggesting that there might be
a size-limited pocket at the para-position of the phenyl
ring at the 4-position and an electron-withdrawing
group is preferred over an electron-donating group
with the ester and amino groups at the 3-, 7-positions.
In general, the activities against HCT-116 of the four
series decreased in descending order: VII, III, VIII,
V .
(IC50 of 2.9 μg mL−1, 5.4 μg mL−1, 5.6 μg mL−1
,
8.4 μg mL−1, 9.6 μg mL−1, respectively), and 4-
chlorophenyl VIIIc, 4-fluorophenyl VIIIb, phenyl VI-
IIa, 4-methylphenyl VIIIe, 4-nitrophenyl VIIIg, 4-
methoxyphenyl VIIIf analogues for the fourth se-
Concerning the activity against HepG-2, all the
series of compounds VIIe, VIIc, IIIg, IIIa (IC50
=
ries with (IC50 of 8.5 μg mL−1, 22.5 μg mL−1
22.6 μg mL−1, 27.0 μg mL−1, 35.2 μg mL−1
,
,
2.5–4.1 μg mL−1) exhibited higher anti-tumour ac-
tivities against HepG-2 than vinblastine (IC50 = 4.6
μg mL−1) and colchicine (IC50 = 10.6 μg mL−1),
39.4 μg mL−1, respectively), exhibited good activ-
ity against HCT-116 in comparison with colchicine
(IC50 = 42.8 μg mL−1). This potency could be at-
tributed to the presence of the fluoro, bromo and
chloro atoms (electron-withdrawing), methyl group
(electron-donating) at the para-position of the phenyl
ring at the 4-position or the phenyl group (electron-
donating) at the 4-position, suggesting that there
might be a size-limited pocket at the para-position
of the phenyl ring at the 4-position for the third se-
ries, or the chloro, fluoro and nitro groups (electron-
withdrawing), phenyl group (electron-donating), me-
thyl and methoxy groups (electron-donating) at the
while compounds VIIb, VIIa, VIIIf and VIIIe (IC50
=
5.6–10.4 μg mL−1) exhibited good activity in com-
parison with colchicine and the other derivatives and
compound IIId (IC50 = 10.6 μg mL−1) was equipo-
tent as colchicine. This was due to the presence of
the substituent, methyl group (electron-donating) or
the chloro atom (electron-withdrawing) for the third
series or nitro group (electron-withdrawing) or the un-
substituted phenyl group (electron-donating) for the
first series at the 4-position. In general, the activities
against HepG-2 of the four series decreased in the de-
scending order: VII, III, VIII, V .

A. M. El-Agrody et al./Chemical Papers 70 (9) 1279–1292 (2016)
1291
Conclusions
& Fun, H. K. (2012). 2-Amino-4-(4-fluorophenyl)-6-methoxy-
4H-benzo[h]chromene-3-carbnitrile. Acta Crystallographica
Section E, 68, o1934–o1935. DOI: 10.1107/s1600536812023
021.
The current interest in the synthesis of 4H-
benzo[h]chromene derivatives is to focus on their anti-
tumour activities as part of a recent research line seek-
ing the development of new heterocyclic compounds as
potent anti-tumour agents (Abd-El-Aziz et al., 2004;
El-Agrody et al., 2011, 2013, 2014a, 2014b; El-Agrody
& Al-Ghamdi, 2011; Sabry et al., 2011; Al-Ghamdi et
al., 2012). Accordingly, in the present study the syn-
thesis of some 2-amino-4H-benzo[h]chromene and 2,7-
diamino-4H-benzo[h]chromene derivatives was con-
ducted, followed by an anti-tumour evaluation of
all the synthesised compounds. Thirty compounds of
2-amino-4H-benzo[h]chromene and 2,7-diamino-4H-
benzo[h]chromene derivatives were prepared and their
structures were elucidated on the basis of IR, 1H
NMR, 13C NMR, 13C NMR-DEPT/APT and MS
data. Compounds VIIIc, VIId, VIIb, VIIe and VIIIg,
respectively, inhibited the growth of the MCF-7 can-
cer cell in comparison with vinblastine and colchicines;
VIIc, VIIIe, Vf and IIIa, respectively, inhibited the
growth of the MCF-7 cancer cell in comparison with
colchicine, while compounds VIIb, VIId, VIIe, IIIa,
VIIa, VIIIc, VIIc, IIId, IIIg, IIIf, IIIb, IIIh, VIIIb,
VIIIa, VIIIe, IIIc, Vg, IIIe, VIIIg, Vf and VIIIf, re-
spectively, inhibited the growth of HCT-116 cancer
cell in comparison with colchicine. In addition, com-
pounds VIIe, IIIg, IIIa and VIIc, respectively, inhib-
ited the growth of the HepG-2 cancer cell in compari-
son with vinblastine and colchicine, while compounds
VIIb, VIIa, VIIIf and VIIIe, respectively, inhibited
the growth of the HepG-2 cancer cell in comparison
with colchicine and the remaining compounds exhib-
ited near or moderate to lower activities in comparison
with the standard drugs vinblastine and colchicine.
A more extensive study is required to determine ad-
ditional anti-tumour parameters in order to gain a
deeper insight into its structure–activity relationship
and to optimise the effectiveness of this series of
molecules, which can then be used in drug design or
the development of anti-tumour therapeutics.
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DOI: 10.1590/s0103-50532010000600010.
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Cole, S. T., Tillequin, F., & Janin, Y. L. (2009). Diversity-
oriented synthesis of furo[3,2-f]chromanes with antimycobac-
terial activity. European Journal of Medicinal Chemistry, 44,
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Bru¨hlmann, C., Ooms, F., Carrupt, P. A., Testa, B., Catto, M.,
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El-Agrody, A. M., Al-Omar, M. A., Amr, A. G. E., Chia, T.,
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El-Agrody, A. M., Abd-Rabboh, H. S. M., & Al-Ghamdi, A. M.
(2013). Synthesis, antitumor activity and structure–activity
relationship of some 4H-pyrano[3,2-h]quinoline and 7H-
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& Al-Dies, A. A. M.
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d]pyrimidine derivatives and structure–activity relationships
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Acknowledgements. This research was supported by a pro-
gramme to support research and researchers at King Khalid
University, Abha, Saudi Arabia, no. KKU-SCI-11-028. The
authors wish to thank the Regional Centre for Mycology &
Biotechnology (RCMP), Al-Azhar University for carrying out
El-Agrody, A. M., Khattab, E. S. A. E. H.,
& Fouda,
A. M. (2014b). Synthesis, structure–activity relationship
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the anti-tumour study and Mr. Ali Y. A. Alshahrani for the
13
preparation of the samples for 1H NMR and
yses.
C NMR anal-
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